Identification of the horse epidermal growth factor (EGF) coding sequence and its use in monitoring EGF gene expression in the endometrium of the pregnant mare.
Abstract: The PCR technique and highly degenerate oligonucleotide primers were used to amplify a 282 bp fragment of the horse (Equus caballus) epidermal growth factor (EGF) cDNA. The clone corresponded to 94 amino acids of the EGF precursor molecule. The deduced amino acid sequence of the 53 residue EGF mitogenic peptide within the precursor sequence showed 60-70% identity with five other published EGF sequences. The PCR cDNA fragment hybridized to a 4.9 kb transcript in horse kidney and endometrial RNA which was of a similar size to the mature EGF transcript found in other mammalian species. The horse cDNA clone was used in Northern blots to monitor EGF expression in the endometrium of pregnant mares up to day 83 of gestation (term = 330-340 days). The level of expression increased from day 33 and showed a further dramatic increase between days 35 and 45, which coincides with the onset of implantation and placentation in this species. Levels remained elevated up to day 83. The horse DNA sequence was used to design sense and antisense oligonucleotide probes (45-mers) for in situ hybridization studies. The antisense probe showed specific hybridization to the glandular, but not lumenal, epithelial cells of the endometrium and there was no signal in fetal membranes. The in situ hybridization signal increased between days 35 and 45 to a similar degree to that observed in the Northern blot analysis. This dramatic increase in EGF expression in the glandular epithelium of the mare's endometrium during pregnancy may provide a mitogenic stimulus to the endometrium and/or trophoblast to facilitate placental differentiation and attachment. Alternatively, the precursor could be involved in the endometrial gland secretory process which is necessary to produce uterine milk for fetal sustenance. The PCR cloning methods used in this study should be generally applicable to the cloning of EGF cDNAs from other species.
Publication Date: 1994-06-01 PubMed ID: 7916972DOI: 10.1677/jme.0.0120341Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research paper focuses on the identification of the horse epidermal growth factor’s (EGF) coding sequence, which assists in monitoring the EGF gene expression in the endometrium of pregnant horses during the gestation period.
Methodology
- The researchers used the polymerase chain reaction (PCR) method and degenerate oligonucleotide primers to amplify a 282 bp fragment of the horse epidermal growth factor (EGF) cDNA.
- The deduced amino acid sequence from the EGF precursor molecule corresponded to 94 amino acids.
- The sequence showed 60-70% identity with five other published EGF sequences.
- The horse cDNA clone was used in Northern blots to monitor EGF expression in the endometrium of pregnant mares up to day 83 of gestation.
Findings
- The amplified PCR cDNA fragment was found to hybridize to a 4.9 kb transcript in horse kidney and endometrial RNA, similar in size to the mature EGF transcript in other mammalian species.
- The EGF expression level increased from day 33 and had a further dramatic increase between days 35 and 45, coinciding with the onset of implantation and placentation in horses. The levels remained elevated up to day 83.
- Using the horse DNA sequence, sense and antisense oligonucleotide probes (45-mers) were designed for in situ hybridization studies.
- The antisense probe showed specific hybridization to the glandular, but not lumenal, epithelial cells of the endometrium, and there was no signal in fetal membranes.
Implications
- The research highlights how an increase in EGF expression in the glandular epithelium of the mare’s endometrium during pregnancy may provide a mitogenic stimulus to facilitate placental differentiation and attachment.
- The research also highlights that EGF precursor could be involved in the endometrial gland secretory process, necessary to produce uterine milk for fetal sustenance.
- The PCR cloning methods used in this study show promise for their application in cloning EGF cDNAs from other species.
Cite This Article
APA
Stewart F, Power CA, Lennard SN, Allen WR, Amet L, Edwards RM.
(1994).
Identification of the horse epidermal growth factor (EGF) coding sequence and its use in monitoring EGF gene expression in the endometrium of the pregnant mare.
J Mol Endocrinol, 12(3), 341-350.
https://doi.org/10.1677/jme.0.0120341 Publication
Researcher Affiliations
- TBA Equine Fertility Unit, Mertoun Paddocks, Newmarket, Suffolk, UK.
MeSH Terms
- Amino Acid Sequence
- Animals
- Base Sequence
- Cloning, Molecular
- DNA, Complementary / genetics
- Endometrium / metabolism
- Epidermal Growth Factor / genetics
- Female
- Gene Expression Regulation
- Genes
- Horses / genetics
- Molecular Sequence Data
- Polymerase Chain Reaction
- Pregnancy
- Pregnancy, Animal / genetics
- Pregnancy, Animal / metabolism
- Templates, Genetic
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